Figure 2

(A) HC69.5. HIV (GFP+) cells were treated with 100 μg/ml poly IC and different concentrations of Δ (9)-THC or CBD (0.01 and 1 μM). Cells were incubated overnight and were fixed using the Fixation and Permeabilization Solution Kit (Cat. No. 554715). Flow cytometry analysis was performed to detect the HIV infection by expressing GFP-positive cells. Immortalized human microglia cells (C20) (HIV-; GFP-) were used as untreated and uninfected control. Results were expressed as percentage of positive cells. Experiments were performed at least three times in replicates. GraphPad Prism 5 software was used to graph and conduct statistical analysis. The normality distribution was evaluated by Kolmogorov–Smirnov test, while statistical significance (p ≤ 0.05) was calculated by ANOVA and Dunns as a post-hoc test. (B) HC69.5. HIV (GFP +) cells were treated with different concentrations of Δ (9)-THC or CBD (0.01 and 1 μM). Cells were incubated overnight and were fixed using the Fixation and Permeabilization Solution Kit (Cat. No. 554715). Flow cytometry analysis was performed to detect the HIV infection by expressing GFP-positive cells. Immortalized human microglia cells (C20) (HIV−; GFP−) were used as untreated and uninfected control. Results were expressed as number of positive cells. Experiments were performed at least three times in replicates. GraphPad Prism 5 software was used to graph and conduct statistical analysis. The normality distribution was evaluated by Kolmogorov–Smirnov test, while statistical significance (p ≤ 0.05) was calculated by ANOVA and Dunns as a post-hoc test. (C) LTR gene expression on HC69.5. HIV (GFP+) cells was assessed by RT PCR. H69.5 cells treated with/without 100 μg/ml poly IC, 1 μM concentration of CBD or Δ (9)-THC as figure is showing. After 24 h of treatments, RNAs were extracted, and reverse transcribed followed by real time PCR for LTR gene (HIV LTR Pa03453409_s1; Thermofisher Scientific). GAPDH (Hs99999905_m1) was used as a housekeeping gene. Data represent the means ± standard error of three independent experiments. Graph represents the transcript accumulation index respect to negative control C20. Data were analyzed using GraphPad Prism software. The normality distribution was evaluated by Kolmogorov–Smirnov test, while statistical significance was calculated by ANOVA and Dunns Multiple Comparison Test as a post-hoc test. Differences were considered significant at p ≤ 0.05.